二乙烯亚胺对脊髓灰质炎病毒Sabin株灭活效果以及灭活疫苗添加佐剂的效果观察
发布时间：2018-09-12 09:21
【摘要】： 随着全球消灭脊髓灰质炎(脊灰)目标的临近,疫苗衍生脊灰病毒(Vaccine-derived Poliovirus, VDPV)的流行、疫苗相关麻痹型病例(Vaccine Associated Paralytic Poliomyelitis, VAPP)的发生等对无脊灰状态成果的巩固威胁很大。在消灭脊灰的最后阶段,应尽快消灭脊灰野毒株,并使用脊髓灰质炎灭活疫苗(Inactivated poliovirus vaccine, IPV)替代目前广泛使用的脊髓灰质炎减毒活疫苗(Oral poliovirus vaccine, OPV)来维持无脊灰状态,消除自然界的活病毒。考虑到疫苗生产的生物安全性等因素,WHO鼓励研制Sabin株脊髓灰质炎灭活疫苗(Sabin IPV)。 本论文旨在针对Sabin II型病毒,寻找一种与传统灭活剂-甲醛相比较,对病毒的抗原破坏更小,且更安全的病毒灭活剂；并进一步探索Sabin IPV中添加佐剂对免疫效果产生的影响,探讨Sabin IPV最适剂量和佐剂应用的意义。 本研究第一部分进行了二乙烯亚胺(BEI)对脊髓灰质炎病毒Ⅱ型Sabin株的灭活效果观察。通过在不同温度下采用4种浓度的BEI及甲醛对脊髓灰质炎病毒Ⅱ型Sabin株进行灭活,测定活病毒含量和D抗原含量,并进行病毒灭活验证试验。结果在37℃条件下,0.002 mol/L浓度的BEI能在48h内完全灭活病毒,灭活后的病毒液D抗原含量回收率为90.6%,显著高于甲醛灭活的回收率(60.6%)。经验证灭活彻底,无活病毒存在。BEI对脊髓灰质炎病毒Ⅱ型Sabin株的D抗原破坏小,能较好的保持疫苗的有效成分。 本研究的第二部分四组不同剂量的Sabin IPV疫苗与佐剂Al(OH)3配伍后免疫Wistar大鼠,通过检测各组大鼠免疫后血清中和抗体水平、特异性IFN-γ的产生情况以及免疫后各组大鼠CD3/CD4/CD8分子表达的差异,以确定使用Al(OH)3佐剂时Sabin IPV的最适剂量。通过微量中和实验检测Sabin株脊髓灰质炎特异性中和抗体发现,疫苗不同剂量产生的中和抗体GMT水平基本是抗原含量高的组高于抗原含量低的组,添加Al(OH)3佐剂组的中和抗体水平也是高于未添加佐剂组的中和抗体水平。ELISPOT测定针对Sabin株脊髓灰质炎病毒的特异性IFN-γ的结果显示,添加佐剂实验组产生的斑点要多于未添加佐剂组,2-①实验组三个型别所产生的斑点数都最多。流式细胞仪检测各组大鼠CD3/CD4/CD8分子表达差异的结果显示,1-①实验组CD3+CD4+/CD3+CD8+比值最高。综上所述,添加佐剂对三个型别的SabinIPV的免疫反应有增强的效果；综合实验结果可认为Ⅰ型采用30 DU/dose,Ⅱ型采用16 DU/dose,Ⅲ型22.5 DU/dose剂量时较为适宜。
[Abstract]:As the global goal of polio eradication approaches, the prevalence of vaccine-derived poliovirus (Vaccine-derived Poliovirus, VDPV) and the occurrence of vaccine-associated paralytic cases (Vaccine Associated Paralytic Poliomyelitis, VAPP) pose a great threat to the consolidation of poliomyelitis (poliomyelitis) status. In the final stage of polio eradication, the polio field strain should be eliminated as soon as possible, and poliomyelitis inactivated vaccine (Inactivated poliovirus vaccine, IPV) should be used to replace the widely used live attenuated poliomyelitis vaccine (Oral poliovirus vaccine, OPV) to maintain poliomyelitis free. Eliminate live viruses from nature. Who encourages the development of Sabin strain inactivated poliomyelitis vaccine (Sabin IPV). Considering the biological safety of vaccine production. The purpose of this paper is to search for a virus inactivated agent with less antigenic damage and more safety to Sabin II virus than traditional inactivated agent-formaldehyde, and to explore the effect of adjuvant added in Sabin IPV on the immune effect. To explore the significance of optimal dosage and adjuvant application of Sabin IPV. In the first part of this study, the inactivation effect of diethylenimide (BEI) on poliovirus type 鈪，
本文编号：2238579
[Abstract]:As the global goal of polio eradication approaches, the prevalence of vaccine-derived poliovirus (Vaccine-derived Poliovirus, VDPV) and the occurrence of vaccine-associated paralytic cases (Vaccine Associated Paralytic Poliomyelitis, VAPP) pose a great threat to the consolidation of poliomyelitis (poliomyelitis) status. In the final stage of polio eradication, the polio field strain should be eliminated as soon as possible, and poliomyelitis inactivated vaccine (Inactivated poliovirus vaccine, IPV) should be used to replace the widely used live attenuated poliomyelitis vaccine (Oral poliovirus vaccine, OPV) to maintain poliomyelitis free. Eliminate live viruses from nature. Who encourages the development of Sabin strain inactivated poliomyelitis vaccine (Sabin IPV). Considering the biological safety of vaccine production. The purpose of this paper is to search for a virus inactivated agent with less antigenic damage and more safety to Sabin II virus than traditional inactivated agent-formaldehyde, and to explore the effect of adjuvant added in Sabin IPV on the immune effect. To explore the significance of optimal dosage and adjuvant application of Sabin IPV. In the first part of this study, the inactivation effect of diethylenimide (BEI) on poliovirus type 鈪，
本文编号：2238579
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